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Calpain-10 (EC 3.4.22.-) (Calcium-activated neutral proteinase 10) (CANP 10) [KIAA1845]


Cloning, expression, and polymorphism of the porcine calpain10 gene.

Calpains are calcium-regulated proteases involved in cellular functions that include muscle proteolysis both ante- and post-mortem. This study was designed to clone the complete coding sequence of the porcine calpain10 gene, CAPN10, to analyze its expression characteristics and to investigate its polymorphism. Two isoforms of the CAPN10 gene, CAPN10A and CAPN10B, were obtained by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends methods combined with in silico cloning. RT-PCR results indicated that CAPN10 mRNA was ubiquitously expressed in all tissues examined and, with increasing age, the expression level increased in muscles at six different growth points. In the same tissues, the expression level of CAPN10A was higher than that of CAPN10B. In addition, three single nucleotide polymorphisms were detected by the PCR-single-stranded conformational polymorphism method and by comparing the sequences of Chinese Min pigs with those of Yorkshire pigs. C527T mutation was a missense mutation and led to transforming Pro into Leu at the 176 th amino acid. The results of the current study provided basic molecular information for further study of the function of the porcine CAPN10 gene.

MeSH Terms

  • Aging
  • Animals
  • Base Sequence
  • Calpain
  • Cloning, Molecular
  • DNA Mutational Analysis
  • Molecular Sequence Data
  • Organ Specificity
  • Polymorphism, Single Nucleotide
  • Swine
  • Swine, Miniature
  • Tissue Distribution

Genetic and nongenetic regulation of CAPN10 mRNA expression in skeletal muscle.

The gene encoding calpain-10 (CAPN10) has been identified as a candidate gene for type 2 diabetes. Our aim was to study the impact of genetic (heritability and polymorphisms) and nongenetic (insulin, free fatty acids, and age) factors on CAPN10 mRNA expression in skeletal muscle using two different study designs. Muscle biopsies were obtained before and after hyperinsulinemic-euglycemic clamps from 166 young and elderly monozygotic and dizygotic twins as well as from 15 subjects with normal (NGT) or impaired glucose tolerance (IGT) exposed to an Intralipid infusion. We found hereditary effects on both basal and insulin-exposed CAPN10 mRNA expression. Carriers of the type 2 diabetes-associated single nucleotide polymorphism (SNP)-43 G/G genotype had reduced CAPN10 mRNA levels compared with subjects carrying the SNP-43 A-allele. Age had no significant influence on CAPN10 mRNA levels. Insulin had no significant effect on CAPN10 mRNA levels, neither in the twins nor in the basal state of the Intralipid study. However, after a 24-h infusion of Intralipid, we noted a significant increase in CAPN10 mRNA in response to insulin in subjects with NGT but not in subjects with IGT. In conclusion, we provide evidence that mRNA expression of CAPN10 in skeletal muscle is under genetic control. Glucose-tolerant but not glucose-intolerant individuals upregulate their CAPN10 mRNA levels in response to prolonged exposure to fat.

MeSH Terms

  • Adult
  • Aged
  • Aging
  • Biopsy
  • Blood Glucose
  • Calpain
  • Fat Emulsions, Intravenous
  • Fatty Acids, Nonesterified
  • Gene Expression Regulation
  • Genotype
  • Glucose Clamp Technique
  • Glucose Intolerance
  • Humans
  • Insulin
  • Middle Aged
  • Muscle, Skeletal
  • Polymorphism, Genetic
  • Polymorphism, Single Nucleotide
  • RNA, Messenger
  • Twin Studies as Topic
  • Twins, Dizygotic
  • Twins, Monozygotic